Abstract of Study (Max 300 Words): Objective: The objective of the present work was to establish the gene knockdown system (RNAi)
using different Multiple Myeloma Cell lines (MMCL).
Material and Methods: RNAi is generally used in biomedical research to study the outcome of
knockdown of gene expression. In this research, retroviral-shRNA expression constructs based onmicro RNA-adapted short hairpin RNA (sh RNA mir) were designed and constructed. The sequenceof the cloned fragments in the vector constructs was determined, and the effect of constructs inMMCLs was examined using qRT-PCR and Western blot analysis.
Results: In this work, we established a method for making retroviral-shRNA expression constructsbased on micro RNA-adapted short hairpin RNA (sh RNA mir) design. The HGF producing human myeloma cell line JJN-3 transduced with lentiviral HGF-shRNA transduction particles wereanalyzed for their effect on the HGF gene expression. HGF knockdown MMCL was analyzedboth at mRNA level and protein levels. Two out of five clones of myeloma cell line JJN-3 (JJN3-sh3308 and JJN3-sh47137) were transduced with lentiviral HGF–shRNA transduction particleswere showing 50-60 % knockdown in their HGF mRNA and protein expression.
Conclusion: In conclusion, the HGF knockdown cell lines can be used as tissue culture models
to investigate the role of HGF protein in the pathology of multiple myeloma. Further, the retro-
viral-HGF-shRNA expression constructs can study HGF response in various multiple myeloma
via gene knockdown studies.
Keywords: Multiple myeloma, Hepatocyte growth factor (HGF), Gene knockdown, short hairpin
RNA
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